一株好氧反硝化细菌的分离鉴定及反硝化能力

利用驯化活性污泥的方法筛选出一株好氧反硝化细菌G3,该菌在碳源充足且溶解氧为(4.0±1.0)mg·L-1条件下,24h内对NO-2-N积累量为12.03 mg·L-1,对NO-3-N的去除率为91.09%.扫描电镜观察其形态特征为(0.4～0.6)μm×(1.2～2.0)μm的短杆菌.16SrDNA序列分析表明,该菌与Pseudomonas stutzeri的相似性为100%.利用MEGA软件对该菌株与已报道的相关微生物进行系统发育分析,并克隆出与好氧反硝化相关的周质硝酸盐还原酶的亚基napA基因,初步判定筛选菌株具有周质硝酸盐还原酶.     

高温好氧反硝化菌的分离鉴定及其反硝化性能研究

通过高温筛选,从燃煤电厂生物滴滤系统填料的生物膜上分离出1株高效好氧反硝化细菌TAD1.该菌株革兰氏染色呈阴性,短杆状,大小为(0.67～0.89)μm×(1.03～1.41)μm;经生理生化特性和基于16S rDNA序列分析,初步鉴定该菌为螯台球菌属(Chelatococcus 8p.).对菌株TAD1的反硝化性能进...     

基于微电极技术的反硝化滤池生物膜特性分析

为研究反硝化滤池中溶解氧对反硝化作用的影响,制备性能良好尖端直径在30μm以内的氧(O2)以及硝酸盐(NO3-)微电极,以此为测试工具,对反硝化滤池中生物膜内部O2、NO3-微环境分布进行测试,通过建立扩散-反应方程,获得生物膜微环境耗氧及反硝化活性特征.研究结果表明,溶解氧在生物膜内部呈明显的下降趋势,从主体溶液氧浓度约1mg/L下降至生物膜300μm深度处约为0.生物膜内部反硝化活性区域发生在300~600μm深度范围内.该条件下反硝化滤池生物膜的氧利用速率常数以及反硝化速率常数之间的比值为1.46,溶解氧对反硝化过程的影响是显著的.     

碳源变化对降解2,4-D的好氧颗粒污泥性能及形态的影响

在长期运行的序批式生物反应器(SBR)中,考察了以葡萄糖和2,4-二氯苯氧乙酸(2,4-D)为混合碳源培养的好氧颗粒污泥在转换为2,4-D唯一碳源废水后,其形态、结构及对目标污染物去除功能的变化.结果表明,基质转换为2,4-D单一碳源后,好氧颗粒污泥仍保持了对目标污染物高效的去除能力.当进水2,4-D浓度为361～564mg/L,其去除率为99.2%～100%,COD平均去除率达到85.6%.混合碳源向2,4-D单一碳源转换对原好氧颗粒结构产生一定破坏作用,使其发生部分解体,粒径由513μm下降到302μm.但好氧颗粒污泥良好的耐负荷冲击使其保持了颗粒主体,通过一段时间适应调整后能够重新聚集生长,最终获得能够利用2,4-D为唯一碳源生长并具有良好沉降性(SVI20～40mL/g)的好氧颗粒污泥,粒径为489μm.扫描电子显微镜(SEM)观察结果表明,混合碳源转向单一碳源使好氧颗粒生物相丰富度降低.     

含硝氮废水的好氧反硝化处理及其系统微生物群落动态分析

利用好氧反硝化细菌强化生物陶粒反应器处理含硝氮废水,探讨了生物陶粒反应器中好氧反硝化生物脱氮的实现过程,并运用变性梯度凝胶电泳技术(DGGE)对微生物菌群结构稳定性进行了分析.结果表明,在水力负荷0.75 m/h,气水比5:1～10:1,水温15～23℃,进水COD负荷1.92～5.98 kg/(m3·d),硝氮负荷O.60～1.34 kg/(m3·d)的条件下,生物陶粒反应器在稳定运行阶段可以基本实现对硝酸氮的完全去除,对总氮的去除率最高可达95.73%,出水中亚硝酸盐一直保持在较低水平.PCR-DGGE指纹图谱显示,微生物多样性与废水的处理效果出现协同变化的特征.在整个运行阶段,好氧反硝化菌群X31在反应器中稳定存在,并始终是优势菌群.     

数学模拟好氧颗粒污泥的形成及水力剪切强度对颗粒粒径的影响

以生物膜数学模型和活性污泥数学模型为基础,建立好氧颗粒污泥一维数学模型,并模拟营养物质的去除、颗粒粒径变化、反应器周期表现以及好氧颗粒污泥内DO和菌群分布.模拟有机物SS浓度和出水NH4+-N浓度逐渐降低,在大约50 d左右达到稳定,50 d后模拟出水浓度分别<25 mg.L-1和<1.5 mg.L-1.模拟出水NO3--N浓度随着粒径的增加呈现降低趋势.当颗粒粒径由模拟30 d时的1.1 mm增加到100 d时的2.5 mm,颗粒污泥缺氧区面积相应增加,总氮(TN)的去除率由不到10%增加到91%左右,最终模拟出水NO3--N浓度降低到<3 mg.L-1.在好氧颗粒污泥系统内,由于氧气传质阻力,模拟颗粒污泥外层DO浓度高而内层浓度低,颗粒内可以发生同时硝化反硝化,且好氧颗粒污泥内DO特征随时间而发生变化.在好氧初期,好氧颗粒污泥代谢活性高,模拟DO传质深度大约为100～200μm;而在好氧末期,模拟DO传质深度为800μm.模拟自养菌主要分布在DO浓度高的颗粒外层,异养菌分布在整个颗粒.当水力剪切系数kde由0.25(m.d)-1逐渐增加到5(m.d)-1时,模拟颗粒平衡粒径依次由3.5 mm左右减小到大约1.8 mm.在不同水力剪切强度下模拟颗粒污泥生长特征相似,其平衡状态下粒径随曝气强度的增加而减小,可以通过控制曝气强度来控制好氧颗粒污泥的平衡粒径.     

生物滤池系统内生化作用机理综合研究

采用淹没式生物滤池系统对官厅水库入库水进行了生物预处理的可行性研究 .综合考察了生物滤池系统内的生化作用机理 ,对地表水中的好氧反硝化进行了探索性研究 ,并在水力负荷为 4m3 (m2 ·h)的情况下取得了 2 9%的脱氮率 .好氧反硝化对于低C N比的地表水进行生物脱氮提供了一种新的思路     

好氧颗粒污泥的培养及其微观特性表征

采用厌氧颗粒污泥为接种污泥,在SBR反应器内培养好氧颗粒污泥,并对其微观特征进行了研究。COD负荷为1.5kg/m3·h～1.8kg/m3·h,表面气体流速在0.0052m/s之间,沉淀时间控制在10～8min时,有利于好氧颗粒污泥的形成。20d后完成好氧颗粒污泥的驯化和培养。研究发现培养的好氧颗粒污泥微生物相以杆菌和丝状菌为主,球菌较少。用扫描电子显微镜(SEM)观察好氧颗粒污泥的微观结构,颗粒污泥具有不平整的表面,轮廓清晰,表面有薄层粘液覆盖并有绒毛状结构。颗粒污泥表面和内部有明显的孔洞或孔隙。研究结果表明,好氧颗粒污泥具有良好的有机物降解能力和同步硝化反硝化能力。     

前置反硝化-硝化生物滤池工艺物料平衡分析

针对前置反硝化(DN)-硝化(CN)生物滤池工艺进行碳(COD)、氮、磷平衡分析,建立了该工艺的物料衡算方法。分析结果表明:DN池中反硝化的碳源主要是甲醇,占DN池反硝化碳源量的70.8%。入流总氮中有13.7%通过CN池的同步硝化反硝化过程去除;入流COD中有46.4%通过CN池中异养菌的好氧氧化去除;入流磷总量约有75.9%被微生物利用。CN池中硝化过程耗氧量为29.0%,占供氧量的比例最大。     

厌氧-好氧-缺氧短程硝化同步反硝化除磷工艺研究

构建了主要由厌氧-好氧-缺氧构成的短程硝化同步反硝化除磷工艺,并在常温条件下用于生活污水的处理.研究发现,通过调节反应器内好氧区的pH(8.2～8.7)和溶解氧(DO为3～5mg·L-1)能实现该工艺的快速启动,在好氧区内实现亚硝酸盐的累积.在稳定运行期内,DO是影响短程硝化的主要影响因素,好氧1区DO控制在1.5～2.0mg·L-1,好氧2区DO控制在0.5～1.0mg·L-1,好氧区内亚硝酸盐氮累积浓度稳定在5～10mg·L-1,氨氮去除率达到90%以上.各反应单元内碳源、硝酸盐和亚硝酸盐对除磷贡献的研究表明,该工艺的缺氧段实现了在不外加碳源的情况下以亚硝酸盐和硝酸盐共同作为电子受体的反硝化除磷,反硝化除磷量占系统总除磷量的80%以上.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in the soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.     

Intraspecific differences in effects of co-contamination of cadmium and

A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p ＜ 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.